Available images

Simple Western: Integrin alpha M/CD11b Antibody [ABIN4099304] - Simple Western lane view shows a specific band for Cd11b in 1.0 mg/ml of Dentate Gyrus from Rat Brain at an antibody concentration of 1:50. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Image provided through customer review.

Application Details

Western Blot 2 μg/mL, Simple Western 1:50, Flow Cytometry 1:10-1:1000, Immunohistochemistry 1:400, Immunocytochemistry/Immunofluorescence 1:200, Immunohistochemistry-Paraffin 1:400, Immunohistochemistry-FrozenThis CD11b antibody is useful for Western blot analysis, Immunocytochemistry, Flow Cytometry (PMID 21422470) and IHC-paraffin embedded sections. In Western blot a specific band is observed ~ 160 kDa and an apparant non-specific band is observed ~ 56 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer ( pH 6.0) is recommended. In ICC/IF, membrane staining was observed in Raw 264.7 cells. This antibody does not appear to work in human with Western blot. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 23980916) The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol

Western Blot protocol specific for CD11b Antibody Western Blot Protocol1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. . Rinse the blot.. Block the membrane using standard blocking buffer for at least 1 hour.. Wash the membrane in wash buffer three times for 10 minutes each.. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.. Wash the membrane in wash buffer three times for 10 minutes each.. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).. Apply the detection reagent of choice in accordance with the manufacturers instructions.**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry-Paraffin Embedded Sections Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:. Wash sections in deionized water three times for 5 minutes each.. Wash sections in wash buffer for 5 minutes. . Block each section with 100-400 µL blocking solution for 1 hour at room temperature. . Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4C. . Remove antibody solution and wash sections in wash buffer three times for 5 minutes each. . Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature. . Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each. . Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature. . Wash sections three times in wash buffer for 5 minutes each. . Add 100-400 µL DAB substrate to each section and monitor staining closely. . As soon as the sections develop, immerse slides in deionized water. . Counterstain sections in hematoxylin. . Wash sections in deionized water two times for 5 minutes each. . Dehydrate sections. . Mount coverslips.

Simple Western: Integrin alpha M/CD11b Antibody [ABIN4099304] - Simple Western lane view shows a specific band for Cd11b in 1.0 mg/ml of Dentate Gyrus from Rat Brain at an antibody concentration of 1:50. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Image provided through customer review.